title: Sample 13
description: Protocols: Peripheral blood mononuclear cells (PBMCs) were isolated from 24 mL peripheral blood of participants by the density gradient centrifugation (Ficoll-Paque PLUS, GE Healthcare, USA). Peripheral blood mononuclear cells (PBMCs) were isolated from 24 mL peripheral blood of participants by the density gradient centrifugation (Ficoll-Paque PLUS, GE Healthcare, USA). PBMCs were differentiated by macrophage colony-stimulating factor (M-CSF) (10ng/ml) (Sigma-Aldrich, USA) in RPMI1640 medium (Gibco, USA) supple-mented with 10% FCS (Gibco, USA) with harvesting after 5 days. Phenotypical maturation of monocyte-derived macrophages was confirmed by light microscopy and flow cytome-try with specific antibodies to CD14+ and CD68+ (eBioscience, USA) RNA was isolated from monocyte-derived macrophages and amplified following the user manual of the SMART-Seq™ v4 Ultra™ Low Input RNA Kit for sequencing. Sequencing libraries were generated using the NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (NEB, Ipswich, MA, USA), following the manufacturer's recommendations.